The Bioassay-Guided Fractionation and Identification of Potent Acetylcholinesterase Inhibitors from Narcissus c.v. ‘Hawera’ Using Optimized Vacuum Liquid Chromatography, High Resolution Mass Spectrometry and Bioautography

Mroczek, Tomasz; Dymek, Aleksandra; Widelski, Jarosław; Wojtanowski, Krzysztof Kamil

doi:10.3390/metabo10100395

Cited by 17 publications

(25 citation statements)

References 31 publications

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“…The aim of this study was to analyze five different solvents and solutions of different polarity as follows: methanol, 1% methanolic tartaric acid, ethyl acetate, dichloromethane and cyclohexane. The plant materials were extracted using PLE in conditions previously outlined and thoroughly tested for other types of the alkaloids in our laboratory [ 29 , 31 , 32 , 33 ]. On the basis of these data, the highest recovery of alkaloids was obtained at elevated temperature of 80 °C and 110 bar pressure.…”

Section: Resultsmentioning

confidence: 99%

“…We used special Oasis HLB sorbent for selective concentration of the active compounds from crude extracts. It has proven its effectiveness for the isolation of other types of the alkaloids studied in our lab [ 31 , 32 ]. Moreover, this method is relatively fast as in short time several extracts were purified.…”

Section: Resultsmentioning

confidence: 99%

“…The working conditions of the Dionex ASE 100 extractor were set as follows: static time 10 min, flush volume 60%, number of extraction cycles 3; for each solvent, the tests were performed twice. This selection of parameters was the result of a number of previous studies conducted in our laboratory [ 29 , 31 , 32 , 33 ]. The obtained extracts were transferred into 50 mL volumetric flasks and diluted to the mark with the solvent used for extraction.…”

Section: Methodsmentioning

confidence: 99%

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Optimization of Pressurized Liquid Extraction of Lycopodiaceae Alkaloids Obtained from Two Lycopodium Species

et al. 2021

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Alkaloids of the Lycopodiaceae family are of great interest to researchers due to their numerous properties and wide applications in medicine. They play a very important role mainly due to their potent antioxidant, antidepressant effects and a reversible ability to inhibit acetylcholinesterase (AChE) enzyme activity. This property is of immense importance due to the growing problem of an increasing number of patients with neurodegenerative diseases in developed countries and a lack of effective and efficient treatment for them. Numerous studies have shown that Lycopodiaceae alkaloids are a rich source of AChE inhibitors. In the obtaining of new therapeutic phytochemicals from plant material, the extraction process and its efficiency is crucial. Therefore, the aim of this work was to optimize the conditions of modern PLE to obtain bioactive alkaloids from two Lycopodium species: L. clavatum L. and L. annotinum L. Five different solvents of different polarity were used for prepared plant extracts in order to compare the alkaloid content in and thereby effectiveness of the entire extraction. PLE parameters were used based on multiple studies conducted that gave the highest alkaloids recovery. Crude extracts were purified using solid-phase extraction (SPE) on Oasis HLB cartridge and examined by HPLC/ESI-QTOF–MS of the highly abundant alkaloids. To the best of our knowledge, this is the first time such high recoveries have been obtained for known Lycopodiaceae alkaloids. The best extraction results of alkaloid-lycopodine were detected in the dichloromethane extract from L. clavatum, where the yield exceeded 45%. The high recovery of annotinine above 40% presented in L. annotinum was noticed in dichloromethane and ethyl acetate extracts. Moreover, chromatograms were obtained with all isolated alkaloids and the best separation and quality of the bands in methanolic extracts. Interestingly, no alkaloid amounts were detected in cyclohexane extracts belonging to the non-polar solvent. These results could be helpful for understanding and optimizing the best conditions for isolating potent AChE inhibitors.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Optimization of Pressurized Liquid Extraction of Lycopodiaceae Alkaloids Obtained from Two Lycopodium Species

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“…To improve the antioxidant effect of the fractions, the column could be eluted with more polar solvents after elution with Hexane-Ethyl Acetate (0:100). Methanol, water, and dilute weak acid solutions allow isolating more polar compounds [53,54].…”

Section: Discussionmentioning

confidence: 99%

Antibacterial Potential by Rupture Membrane and Antioxidant Capacity of Purified Phenolic Fractions of Persea americana Leaf Extract

et al. 2021

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The present research focused on evaluating the antibacterial effect and the mechanism of action of partially purified fractions of an extract of Persea americana. Furthermore, both its antioxidant capacity and composition were evaluated. The extract was fractionated by vacuum liquid chromatography. The antimicrobial effect against Staphylococcus aureus (ATCC 6538), Escherichia coli (ATCC 11229), Pseudomonas aeruginosa (ATCC 15442), and Salmonella choleraesuis (ATCC 1070) was analyzed by microdilution and the mechanism of action by the Sytox green method. The antioxidant capacity was determined by DPPH, FRAP, and ABTS techniques and the composition by Rp-HPLC-MS. All fractions showed a concentration-dependent antibacterial effect. Fractions F3, F4, and F5 (1000 µg/mL) showed a better antibacterial effect than the extract against the bacteria mentioned. The F3 fraction showed inhibition of 95.43 ± 3.04% on S. aureus, F4 showed 93.30 ± 0.52% on E. coli, and F5 showed 88.63 ± 1.15% on S. choleraesuis and 86.46 ± 3.20% on P. aeruginosa. The most susceptible strain to the treatment with the extract was S. aureus. Therefore, in this strain, the bacterial membrane damage induced by the extract and fractions was evidenced by light fluorescence microscopy. Furthermore, the extract had better antioxidant action than each fraction. Finally, sinensitin was detected in F3 and cinnamoyl glucose, caffeoyl tartaric acid, and cyanidin 3-O-(6′′-malonyl-3′′-glucosyl-glucoside) were detected in F4; esculin and kaempferide, detected in F5, could be associated with the antibacterial and antioxidant effect.

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“…GC‐MS analysis of in vitro cultures from this cultivar indicated the presence of 6‐ epi ‐mesembranol, mesembrenol, 6‐ epi ‐mesembrenol, mesembrine, mesembrenone, lycorine, and haemanthamine 15 . HPLC/ESI‐QTOF‐MS analysis of fractions from intact plants resulted in the detection of one Sceletium ‐type alkaloid mesembrinol (mesembranol) and other 12 Amaryllidaceae alkaloids, including some unexpected compounds, based only on high mass accuracy CID spectra and interpretation of their fragmentation pathways 16 …”

Section: Introductionmentioning

confidence: 99%

GC‐MS analysis of Amaryllidaceae and Sceletium‐type alkaloids in bioactive fractions from Narcissus cv. Hawera

Berkov

Pechlivanova

Denev

et al. 2021

Rapid Comm Mass Spectrometry

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Rationale Narcissus cv. Hawera has been found to biosynthesize some Sceletium‐type alkaloids with antidepressant and anxiolytic activities. This ornamental plant has been poorly studied as a source of bioactive alkaloids including some contraversive reports on in vitro and intact plants. In this study, a detailed GC‐MS characterization of its alkaloid fractions is presented. Methods GC‐MS was used for the identification of compounds in the alkaloid fractions. Both underivatized and silylated samples were analyzed simultaneously. Elevated plus maze and tail suspension tests were used to assay the anxiolytic and antidepressant activities. Ellman’s and MTT‐dye reduction assays were used to evaluate the acetylcholinesterase (AChE) inhibitory and cytotoxicity activities, respectively. Results Of the 29 alkaloids, 13 of Sceletium‐type were detected. Two new alkaloids were identified as 2‐oxo‐mesembrine and 2‐oxo‐epi‐mesembrenol. Lycorine was found as a major compound (43.5%) in the crude silylated methanol extract. After the elimination of lycorine by pre‐crystallization, the major alkaloids were 40.8% 6‐epi‐mesembranol, 16.2% 6‐epi‐mesembrenol, and 13.8% sanguinine. This fraction showed anxiolytic and antidepressant‐like activities as well as potent AChE inhibitory and antineoplastic activities. Conclusions Silylation of the alkaloid fractions from Narcissus cv. Hawera provides better separation, structural information, and improved sensitivity for compounds with two and more hydroxyl groups. The lycorine‐free alkaloid fraction shows a great potential for further pharmacological studies.

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The Bioassay-Guided Fractionation and Identification of Potent Acetylcholinesterase Inhibitors from Narcissus c.v. ‘Hawera’ Using Optimized Vacuum Liquid Chromatography, High Resolution Mass Spectrometry and Bioautography

Cited by 17 publications

References 31 publications

Optimization of Pressurized Liquid Extraction of Lycopodiaceae Alkaloids Obtained from Two Lycopodium Species

Optimization of Pressurized Liquid Extraction of Lycopodiaceae Alkaloids Obtained from Two Lycopodium Species

Antibacterial Potential by Rupture Membrane and Antioxidant Capacity of Purified Phenolic Fractions of Persea americana Leaf Extract

GC‐MS analysis of Amaryllidaceae and Sceletium‐type alkaloids in bioactive fractions from Narcissus cv. Hawera

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